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Telomere-Driven Diseases and Telomere-Targeting Therapies

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Telomere-Driven Diseases and Telomere-Targeting Therapies JCB: Review Telomere-driven diseases and telomere-targeting therapies Paula Martínez and Maria A. Blasco Telomeres and Telomerase Group, Molecular Oncology Program, Spanish National Cancer Centre, Madrid E-28029, Spain Telomeres, the protective ends of linear chromosomes, structure has been proposed to protect chromosome ends from shorten throughout an individual’s lifetime. Telomere degradation and DNA repair activities as well as from telomer- ase activity (Fig. 1, A and B; Griffith et al., 1999; Doksani et al., shortening is proposed to be a primary molecular cause 2013). Telomeres are bound by a specialized complex known of aging. Short telomeres block the proliferative capacity as shelterin that has crucial functions in telomere length regu- of stem cells, affecting their potential to regenerate tissues, lation and in the protection of telomeres from the DNA dam- and trigger the development of age-associated diseases. age response (DDR) by masking the chromosome ends from the DNA repair machinery through repression of the ATM and Mutations in telomere maintenance genes are associated ATR signaling pathways (Palm and de Lange, 2008; Fig. 1 C). with pathologies referred to as telomere syndromes, in- The shelterin complex is composed of six proteins: telomeric cluding Hoyeraal-Hreidarsson syndrome, dyskeratosis repeat binding factors 1 and 2 (TRF1 and TRF2), TRF1- interacting protein 2 (TIN2), protection of telomeres protein 1 congenita, pulmonary fibrosis, aplastic anemia, and liver (POT1), TIN2, POT1-interacting protein (TPP1), and repressor/ fibrosis. Telomere shortening induces chromosomal insta- activator protein 1 (RAP1; Fig. 1 C; de Lange, 2005; Martínez bility that, in the absence of functional tumor suppressor and Blasco, 2010, 2011). genes, can contribute to tumorigenesis. In addition, muta- Telomeres shorten with each cell division as a result of the incomplete replication of linear DNA molecules by conven- tions in telomere length maintenance genes and in shel- tional DNA polymerases, which is called the end-replication terin components, the protein complex that protects problem (Watson, 1972; Olovnikov, 1973). Telomerase com- telomeres, have been found to be associated with differ- pensates for telomere attrition through de novo addition of TTA ent types of cancer. These observations have encouraged GGG repeats onto chromosome ends in those cells where it is normally expressed, such as pluripotent stem cells and adult the development of therapeutic strategies to treat and pre- stem cell compartments (Liu et al., 2007; Flores et al., 2008; vent telomere-associated diseases, namely aging-related Marion et al., 2009). Telomerase is composed of a reverse tran- diseases, including cancer. Here we review the molecular scriptase subunit (TERT) as well as an associated RNA compo- nent (Terc), which is used as a template for the de novo addition mechanisms underlying telomere-driven diseases and of telomeric repeats (Fig. 1 C; Greider and Blackburn, 1985). highlight recent advances in the preclinical development Although telomerase is expressed in adult stem cell compart- of telomere-targeted therapies using mouse models. ments, this is not sufficient to counteract telomere attrition associated with cell division throughout life, and therefore telo- Telomeres, telomerase, and shelterins meres shorten with age in vitro and in vivo (Harley et al., 1990; THE JOURNAL OF CELL BIOLOGY CELL OF JOURNAL THE Telomeres form a special heterochromatic structure at the end Hastie et al., 1990; Lindsey et al., 1991; Collado et al., 2007; of linear chromosomes that protects them from degradation and Liu et al., 2007; Flores et al., 2008; Marion et al., 2009). This DNA repair and recombination activities. Thus, telomeres are progressive telomere shortening eventually leads to critically essential to ensure chromosome stability (Blasco, 2005; Palm short telomeres that can impair the regenerative capacity of tis- and de Lange, 2008). Mammalian telomeres comprise several sues and has been proposed as one of the molecular hallmarks kilobases, between 10 and 15 kb in humans and 25 and 50 kb in of aging (López-Otín et al., 2013). In mice, it has been shown mice, of tandem TTA GGG DNA repeats (Blasco, 2005). Telo- that the rate of increase in the percentage of short telomeres, meres are characterized by the presence of a 30–400-nucleo- rather than the rate of telomere shortening throughout life, is a tide-long 3′ overhang of a G-rich strand, known as the G-strand significant predictor of life span (Vera et al., 2012). Shortened overhang. The G-strand overhang can fold back and invade the telomeres induce a DDR that leads to a growth arrest, during double-stranded telomeric region, forming the so-called T-loop which cells attempt to repair the damage and, if DNA damage and generating a displacement loop, or D-loop. The T-loop is irreparable, triggers replicative senescence (Zou et al., 2004; Fumagalli et al., 2012). Senescent cells progressively accumu- Correspondence to Maria A. Blasco: [email protected] Abbreviations used: 6-thio-dG, 6-thio-2 -deoxyguanosine; AAV, adeno-associ- ′ © 2017 Martínez and Blasco This article is distributed under the terms of an Attribution– ated vector; ALT, alternative lengthening of telomeres; BM, bone marrow; DC, Noncommercial–Share Alike–No Mirror Sites license for the first six months after the dyskeratosis congenita; DDR, DNA damage response; HHS, Hoyeraal-Hreidars- publication date (see http ://www .rupress .org /terms /). After six months it is available under son syndrome; HSPC, hematopoietic stem/progenitor cell; IPF, idiopathic PF; a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International MC, mitotic catastrophe; PF, pulmonary fibrosis; TPE, telomere position effect. license, as described at https ://creativecommons .org /licenses /by -nc -sa /4 .0 /). The Rockefeller University Press $30.00 J. Cell Biol. Vol. 216 No. 4 875–887 https://doi.org/10.1083/jcb.201610111 JCB 875 Figure 1. The shelterin complex and the structure of telomeres. (A) Representative image of a metaphasic chromosome stained with DAPI (blue) and the telomeric DNA–specific peptide nucleic acid probe (yellow). (B) Schematic model of the shelterin complex bound to a telomere in a T-loop configuration. Telomeres contain a double-stranded region of TTA GGG repeats and a 150–200-nucleotide-long single-stranded DNA overhang of a G-rich strand. The G-strand overhang (gray) invades the telomeric double-stranded DNA region to form a protective T-loop, with a displacement D-loop at the invasion site. (C) Schematic representation of telomere-bound proteins, the shelterin complex, and telomerase. The shelterin complex is composed of the telomeric repeat binding factor 1 (TRF1, also known as TERF1), TRF2 (also known as TERF2), repressor-activator protein 1 (RAP1, also known as TERF2IP1), POT1-TIN2 organizing protein (TPP1, also known as ACD), TIN2 (also known as TIFN2), and protection of telomeres protein 1 (POT1). TRF1, TRF2, and POT1 bind directly to telomeric DNA repeats, with TRF1 and TRF2 binding to telomeric double-stranded DNA and POT1 to the 3′ single-stranded G-overhang. TIN2 binds TRF1 and TRF2 through independent domains and recruits the TPP1–POT1 complex, constituting the bridge among the different shelterin components. Telomerase is a two-partner enzyme, the catalytic subunit (TERT) and the RNA template (TERC), that recognizes the 3′-OH at the end of the G-strand over- hang and elongates the telomere. late during life and secrete factors that influence age-associated Lee et al., 1998; Herrera et al., 1999; Leri et al., 2003; Ferrón et diseases (Tchkonia et al., 2013). Indeed, senescence has been al., 2004). Telomere shortening with age has also been observed proposed as a mechanism that evolved to protect from cancer, in several mouse stem cell compartments from different tissues with the drawback of promoting age-associated diseases (Shay, independently of high or low tissue proliferation rates (Flores 2016). Therapeutic interventions based on either chemical acti- et al., 2008). Telomerase-deficient mice show a worsening of vators of telomerase or telomerase-based gene therapy are cur- phenotypes with increasing mouse generations owing to inher- rently being investigated in mouse models for their potential to ited progressively shorter telomeres, with the later generations improve health and extend life span, and as a treatment for short showing severe phenotypes and premature death at pre-repro- telomere syndromes (Bernardes de Jesus et al., 2011; Bär et al., ductive ages (Lee et al., 1998; Herrera et al., 1999; García-Cao 2014, 2015; Bär and Blasco, 2016). et al., 2002). This phenomenon first indicated genetic antici- pation associated with telomerase deficiency. However, and in Aging-associated diseases and support of critical telomere shortening being a determinant of telomeropathies aging and longevity, increased TERT expression in the context Telomeres shorten progressively throughout an individual’s life- of cancer-resistant transgenic mice was sufficient to delay aging time (Cawthon et al., 2003; López-Otín et al., 2013; Sanders and and extend mouse longevity by 40% (Tomás-Loba et al., 2008). Newman, 2013). Telomere attrition in stem cell compartments More recently, these findings have been translated into a po- impairs their tissue and self-renewal capacity and is considered tential therapeutic strategy by using adeno-associated vectors to be one of the primary molecular causes of aging and the
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